Since earthworms live in hostile conditions, they have well-developed defense mechanisms to fight against the invading microbes. In the present study antibacterial activity of the coelomic fluid of earthworm, Eudrilus eugeniae (EE) was tested against selected Gram positive bacterial pathogens namely, Staphylococcus aureus and Bacillus subtilis procured from MTCC, Chandigarh, India. Two different screening techniques were followed under laboratory conditions to look for efficiency of coelomic fluid in suppressing growth of the pathogens. Coelomic fluid (CF) was collected from earthworm body cavity by cold shock method. In vitro antibacterial assay was done by agar dilution and disc diffusion techniques. The results indicate that S. aureus is the most sensitive organism between the two tested pathogens.  Tetracyclin and Streptomycin standard discs were used for positive control. The level of suppression by CF was almost on par with that of tetracyclin. This preliminary study is an indication that there is a possibility for identifying and using the antibacterial factor from CF of earthworm to suppress these pathogens in place of prevailing antibiotics in the market.